Psychomotor stimulant drugs such as amphetamine produce multiple effects. Notable among them is their ability to activate brain dopamine (DA) neurotransmission, increase locomotor activity and support self-administration in humans and laboratory animals. When repeatedly administered, their ability to produce these effects becomes enhanced so that re-exposure to the drug, weeks to months later, produces greater DA activation, locomotion, and work-output aimed at obtaining the drug. These findings support the proposal that sensitization of the appetitive effects of amphetamine and other psychostimulants promotes the pursuit and self-administration of these drugs and may underlie the transition from casual drug use to compulsive drug taking and abuse. A number of long-lasting neuroadaptations have been identified in forebrain regions like the nucleus accumbens (NAcc) that provide neuronal correlates for the expression of behavioral sensitization by amphetamine. Several lines of converging evidence now indicate a coordinated interaction between DA and glutamate in the expression of amphetamine sensitization, one in which the regulation of AMPA receptors is essential, and that is dependent on pre- and postsynaptic PKC signaling in the NAcc. This proposal builds on new data from the laboratory showing that DA can regulate AMPA receptor trafficking and function to enable the expression of behavioral sensitization. It also incorporates recently published findings showing new mechanisms by which PKC can directly and indirectly regulate AMPA receptor insertion and function. Together, these findings support the hypothesis that NAcc AMPA receptors activate medium spiny neurons and the ensuing motivated behavior the animal displays while Gq-coupled DA receptors enable the expression of sensitization by initiating PKC-mediated signaling to regulate AMPA receptor trafficking and function. To begin testing this hypothesis, the experiments outlined in this proposal will use a model of enhanced amphetamine self-administration and reinstatement to determine the role played by different PKC substrates in the NAcc in the expression of these sensitized behaviors. The proposed experiments exploit a multidisciplinary approach, using behavioral, biochemical, pharmacological, and viral-mediated gene transfer techniques. In Aims 1 and 2, viral-mediated gene transfer will be used to determine the contribution of PKC phosphorylation of GluR1 and neurogranin specifically in NAcc neurons to the expression of sensitization. In Aim 3, pharmacological inhibition of PKC will be used to assess its ability to enable the expression of amphetamine sensitization by regulating presynaptic DA overflow. By deciphering the neuroadaptations that underlie the expression of sensitization, we will increase our understanding of the mechanisms underlying the enhanced pursuit and self- administration of psychostimulant drugs and inform the development of therapeutic strategies aimed at reversing these maladaptive behaviors.